WO2010116912A1

WO2010116912A1 - Tracking device, tracking microscope provided with the tracking device, and tracking method

Info

Publication number: WO2010116912A1
Application number: PCT/JP2010/055514
Authority: WO
Inventors: 正俊石川; 寛雅奥
Original assignee: 国立大学法人東京大学
Priority date: 2009-04-08
Filing date: 2010-03-29
Publication date: 2010-10-14
Also published as: JP5263886B2; JP2010243868A

Abstract

Provided is a tracking device capable of tracking objects of which the images are to be captured without complicating. The visual feedback controller of the tracking device comprises: a detection means for detecting the respective relative movements of the objects of which the images are continuously captured beyond a predetermined period with respect to the preparation; an object extraction means for extracting the object, the relative movements of which are detected, as candidates to be tracked; and a selection means for selecting, as objects to be tracked, the objects selected by a user from among the candidates to be tracked that are extracted and displayed on a display means.

Description

Tracking device, tracking microscope equipped with the tracking device, and tracking method

The present invention relates to a tracking microscope including a tracking device that tracks a captured moving object by performing visual feedback on the image of the captured moving object.

As a device for tracking a moving object, there is a tracking microscope proposed by the present inventors (hereinafter, referred to as “conventional tracking microscope” as appropriate). A conventional tracking microscope is a tracking microscope equipped with a tracking device for tracking (two-dimensional tracking) a moving object (microorganism) by visual feedback (Visual Feedback) (hereinafter referred to as “conventional tracking microscope” as appropriate). (Refer nonpatent literature 1). A general microscope is a very important instrument for observing the movement of microorganisms with sufficient resolution. However, even with such a microscope, there is a problem that it is very difficult to continuously observe the moving microorganisms because the field of view is limited. This is because, when the movement of microorganisms is extremely fast, it is quickly out of view. In order to solve such a problem, the conventional tracking microscope is configured so that the field of view can be moved so that the target microorganism is at the center of the field of view. The movement of the visual field is made possible by a movable stage included in the tracking microscope, and the observation object on the movable stage is brought to the center of the visual field by high-speed movement of the movable stage. (See Non-Patent Document 2 and Non-Patent Document 3). Furthermore, although the conventional tracking microscope performs two-dimensional tracking as described above, three-dimensional tracking is also possible by moving the movable stage and the objective lens up and down (see Non-Patent Documents 4 and 5). In addition to these, Non-Patent Documents 6 and 7 show the configuration of a drive unit for periodically expanding and contracting the shape of a piezoelectric element (piezo element) whose shape changes according to an applied voltage.

(First issue)
However, according to the conventional tracking microscope, the non-moving object looks more stable for the tracking device than the moving object. There was a risk of becoming. Here, for example, consider a case where microorganisms that are moving objects and dust (trash) that are non-moving objects are mixed. In this case, the tracking device at the moment when the microorganism being tracked disappears from the field of view, recognizes dust as a new tracking target instead, and continues the state as it is. A typical example of tracking failure is to track a non-moving object (tracking in a stationary state) without tracking the moving object that is the original target. The provision of a tracking device or a tracking microscope in which a tracking failure is difficult to occur is a first problem to be solved by the present invention.

(Second problem)
As described above, the object to be observed is brought to the center of the visual field by the high-speed movement of the movable stage. Here, it is necessary to realize a state in which the surface on which the movable stage moves first and the preparation surface are parallel (alignment) with micrometer accuracy, but in the past, this state often cannot be realized, With the movement of the movable stage, the position of the preparation surface also moves up and down, and there is a risk that it will deviate from the focal plane. The high-speed movement of the movable stage moves the preparation fixed on it at high speed, and further, the start stop and the direction change for tracking are repeated. Secondly, since one end side of the conventional preparation is cantilevered, the other end side not supported at the time of this change of direction sometimes vibrates up and down due to inertia and elasticity. As a result of this vibration displacing the other end side of the preparation to the point where the depth of field of the microscope has been exceeded, the focus may not be achieved and a clear image may not be obtained. Furthermore, when trying to solve the first and second problems described above, the fixing mechanism for fixing the preparation to the movable stage has a structure that is solid and difficult to attach and detach, and tends to be unusable. In the tracking device or the tracking microscope, it is a second problem to be solved by the present invention to prevent a blurred image caused by a deviation from the focal plane of the preparation or being out of focus while maintaining usability. .

(Third issue)
The movable stage is moved by a tracking device provided in the tracking microscope. However, there is room for improvement in the development and adjustment of the tracking device. Various field experiments are required for the development of the tracking device, but according to the conventional tracking device, it is necessary to prepare an observation object that is actually alive. For example, preparing an observation target such as Paramecium or squirt sperm alive requires enormous effort and cost to create the environment, and may not be collected depending on the season. Furthermore, even if a living observation target can be prepared, the movement direction and speed thereof cannot be controlled, so that there are many cases where it is not suitable as an observation target during preparation. From this background, the tracking microscope could not be prepared as desired, and improvements were desired in this respect. The third problem to be solved by the present invention is to provide a tracking device or a tracking microscope that can be developed and prepared at low cost and under control.

(Fourth issue)
The movement of the movable stage is performed by driving by the tracking device as described in the third problem above. This is because the tracking microscope equipped with the developed and adjusted tracking device is used. The point which should be improved also in. That is, in the conventional tracking microscope, the tracking target cannot be easily changed during observation. For example, when tracking a certain moving object, there may be a case where another moving object in the field of view is desired. When observing a stationary object, there are cases where it is desired to track and observe a moving object and vice versa. In such a case, the conventional tracking microscope cannot easily change the observation target. It is a fourth problem to be solved by the present invention to provide a tracking device or a tracking microscope that can easily change an observation target during observation.

(Fifth issue)
As described in the background art section above, there are two-dimensional and three-dimensional types of tracking in conventional tracking microscopes. Among these, in 3D tracking, the depth position of the target is measured at 1000 Hz by a method called Depth From Diffraction (DFDi), and the movable stage or objective lens is moved up and down (moved in the Z-axis direction) to move in the Z-axis direction. Realize tracking. However, the DFDi method is effective only in combination with a limited number of objects and lighting methods, and has limited versatility. The fifth problem to be solved by the present invention is to provide a tracking device or a tracking microscope capable of solving this problem and realizing three-dimensional tracking with an arbitrary object / illumination technique / observation technique.

The present invention has been made to solve the above-described five problems, and it is difficult to generate tracking failure and unclear images, is easy to develop and adjust, can easily change an observation target, and is further complicated. An object of the present invention is to provide a tracking device, a tracking microscope, and a tracking method that enable three-dimensional tracking.

In order to achieve the above-mentioned object, the present invention has the features described in the following section. Note that the definitions of terms used to describe the characteristics of the invention of any claim are applicable to the inventions of other claims as far as possible, regardless of the order of description or category. Shall be.

(Characteristics of the invention of claim 1)
The tracking device according to the first aspect of the invention (hereinafter referred to as “the device of the first aspect” as appropriate) is a rectangular preparation (chamber) that holds a moving object (for example, a microorganism or a non-moving object may be included). ), A movable stage in which the rectangular preparation is fixed via a fixing mechanism, an objective lens arranged to face the rectangular preparation, and an imaging device that captures an image of a moving object imaged by the objective lens A display device that visually displays an image of the moving object imaged by the imaging device, and a two-dimensional or tertiary representation of the moving object imaged by performing visual feedback on the object image captured by the imaging device. And a control device that controls the movable stage for original tracking. Here, for the imaging target that the control device continues to capture images over a predetermined time, a detection unit that detects individual relative movement with respect to the preparation, and the target for which relative movement is detected by the detection unit. A target extraction unit that extracts an imaging target as a tracking target candidate, and a selection that selects a target to be imaged selected by the user from the tracking target candidates extracted by the target extraction hand and displayed on the display unit. Means.

According to the apparatus of claim 1, the image of the moving object held in the rectangular preparation is picked up by the image pickup device through the objective lens. The moving object is tracked two-dimensionally or three-dimensionally by performing visual feedback on the captured object image. The tracking is realized by a control device (for example, a computer) controlling the movable stage. Here, each object to be imaged that moves relative to the slide is extracted, and the object to be imaged selected from the extracted objects is tracked as a tracking object. The user selects the tracking target while looking at the display means. A captured image usually includes a non-moving object as well as a large number of moving objects, and a tracking target is selected from them. This selection can eliminate non-moving objects and unwanted moving objects. By this elimination, it is possible to effectively eliminate a tracking failure that would otherwise have occurred (first problem).

(Characteristics of the invention described in claim 2)
A tracking device according to claim 2 (hereinafter, referred to as “device of claim 2” as appropriate) indicates the coordinates of each of the imaging targets that the target extraction unit has set as a tracking target candidate on the premise of the device of claim 1. It is configured to grasp target coordinates, and the selection means is configured to grasp point coordinates indicating the coordinates of the point selected by the user on the display means, and the selection means When the point coordinates are grasped, the imaging target related to the coordinates closest to the point coordinates is selected as the tracking target.

According to the apparatus of claim 2, in addition to the operational effects of the apparatus of claim 1, the selection unit compares the coordinates of each of the imaging targets as tracking target candidates with the point coordinates, and the point coordinates are grasped. The target to be imaged according to the coordinates closest to the point coordinates is selected as the tracking target. Each target to be imaged as a tracking candidate is moving relative to the preparation, so that the user's selection may not always be easy. Although the user tries to select a target to be imaged, the timing may not match. In such a case, tracking failure can be effectively suppressed by setting the imaging target closest to the point coordinates as the tracking target (first problem). Used when an imaging target that is different from the target to be selected by the user becomes a tracking target, that is, when an imaging target that is different from the target to be selected is closer to the selection point The person can continue tracking as it is, but can also track the imaging target to be selected by redoing the selection operation.

(Characteristics of Claim 3)
A tracking device according to the invention described in claim 3 (hereinafter referred to as “device of claim 3” as appropriate) includes a rectangular preparation for holding a moving object, and a movable stage in which the rectangular preparation is fixed via a fixing mechanism. An objective lens disposed opposite to the rectangular preparation, an imaging device that captures an image of a moving object imaged by the objective lens, and a display that visually displays the image of the moving object captured by the imaging device And a control device for controlling the movable stage in order to track a moving object imaged two-dimensionally or three-dimensionally by performing visual feedback on the object image imaged by the imaging device. It is a device provided. Here, a calculation unit that calculates an average relative speed with respect to the preparation for the imaging target that is continuously imaged over a predetermined time, and an average relative speed of the imaging target that is calculated by the calculation unit. Are compared with a predetermined reference speed to determine a slow speed, and the imaged target for which the average relative speed is determined to be faster than the reference speed by the determination means is extracted as a tracking target candidate. A target extraction unit; and a selection unit that selects, as a tracking target, a target to be imaged that satisfies a user's selection command or a predetermined selection condition from among tracking target candidates extracted by the target extraction hand. ing.

According to the apparatus of claim 3, the image of the moving object held in the rectangular preparation is picked up by the image pickup device via the objective lens. The moving object is tracked two-dimensionally or three-dimensionally by performing visual feedback on the captured object image. The tracking is realized by the control device controlling the movable stage. Here, an imaged object to be tracked is extracted from an imaged object whose average relative speed is faster than a predetermined reference speed, and the tracking target is selected from the extracted tracking objects. The tracking target is selected depending on either the user's selection command or satisfaction of a predetermined selection condition (for example, the fastest is the selection condition). Usually, a captured image includes a non-moving object as well as a large number of moving objects, and it is necessary to select a tracking target from among them. This is because the non-moving object and the unnecessary moving object are excluded because the non-moving object looks more stable for the tracking device than the moving object, as described above. By this elimination, it is possible to effectively eliminate a tracking failure that would otherwise have occurred (first problem).

(Feature of the invention of claim 4)
A tracking device according to claim 4 (hereinafter, referred to as “apparatus of claim 4” as appropriate) includes a rectangular preparation for holding a moving object, a movable stage in which the rectangular preparation is fixed via a fixing mechanism, and the rectangular The objective lens placed opposite to the preparation, the imaging device that captures the image of the moving object imaged by the objective lens, and the object image captured by the imaging device are captured by visual feedback. And a control device for controlling the movable stage in order to track the moving object two-dimensionally or three-dimensionally. Here, the control device (for example, a computer) uses a target designating unit (for example, a mouse or a keyboard) for arbitrarily designating an imaging target, and the imaging target specified by the target designating unit as a tracking target. Object selection means for selecting.

According to the apparatus of claim 4, the image of the moving object held in the rectangular preparation is picked up by the image pickup device through the objective lens. The moving object is tracked two-dimensionally or three-dimensionally by performing visual feedback on the captured object image. The tracking is realized by the control device controlling the movable stage. Here, the imaging target specified by the user of the apparatus by the input from the target specifying means is selected as a tracking target and is tracked. Usually, a captured image includes a non-moving object as well as a large number of moving objects, and it is necessary to select a tracking target from among them. This is because the non-moving object and the unnecessary moving object are excluded because the non-moving object looks more stable for the tracking device than the moving object, as described above. By this elimination, it is possible to effectively eliminate a tracking failure that would otherwise have occurred (first problem).

(Feature of the invention of claim 5)
A tracking device according to claim 5 (hereinafter, referred to as “apparatus of claim 5” as appropriate) includes a rectangular preparation for holding a moving object, a movable stage in which the rectangular preparation is fixed via a fixing mechanism, and the rectangular The objective lens placed opposite to the preparation, the imaging device that captures the image of the moving object imaged by the objective lens, and the object image captured by the imaging device are captured by visual feedback. And a control device for controlling the movable stage in order to track the moving object two-dimensionally or three-dimensionally. Here, a calculation unit that calculates an average relative speed with respect to the chamber for the imaging target that is continuously imaged over a predetermined time, and an average relative speed of the imaging target calculated by the calculation unit. Are compared with a predetermined reference speed to determine a slow speed, and the imaged target for which the average relative speed is determined to be faster than the reference speed by the determination means is extracted as a tracking target candidate. A target extracting means; and a selecting means for selectively selecting a target to be imaged according to a user's selection instruction or satisfaction of a predetermined selection condition from among tracking target candidates extracted by the target extractor;
Mode switching means for switching between the manual tracking mode and the automatic tracking mode. Further, here, the selection of the selection means is determined in advance according to the user's selection command when the mode switching means is switched to the manual tracking mode, and when the mode switching means is switched to the automatic tracking mode. The imaging target is selected as a tracking target in accordance with satisfaction of the selected condition.

According to the apparatus of claim 5, the image of the moving object held in the rectangular preparation is picked up by the image pickup device via the objective lens. The moving object is tracked two-dimensionally or three-dimensionally by performing visual feedback on the captured object image. The tracking is realized by a control device (for example, a computer) controlling the movable stage. Here, an object to be imaged having an average relative speed higher than a predetermined reference speed is extracted, and tracking is performed by selecting from the extracted tracking objects. The tracking target is selected depending on either the user's selection command or satisfaction of a predetermined selection condition (for example, the fastest is the selection condition). Which is involved depends on the switching of the mode switching means. That is, when the mode switching means is switched to the manual tracking mode, the tracking target is selected based on the user's selection command. Conversely, when the mode switching means is switched to the automatic tracking mode, a predetermined selection condition is satisfied. Is selected for tracking. By enabling the switching of the tracking mode in this way, it becomes possible to select an object in accordance with the user's purpose. Further, by extracting the tracking target, it is possible to effectively exclude non-moving objects and unnecessary moving objects. By this elimination, it is possible to effectively eliminate a tracking failure that would otherwise have occurred (first problem).

(Characteristics of the invention described in claim 6)
In the tracking device according to the sixth aspect (hereinafter referred to as “the device according to the sixth aspect” as appropriate), as a preferable aspect of the device according to any one of the first to fifth aspects, the fixing mechanism has the following configuration. That is, the fixing mechanism includes a fixing portion that is directly fixed to the movable stage and a rectangular frame supported by the fixing portion, and the rectangular frame supports the both ends in the longitudinal direction of the rectangular preparation downward. A pair of supporting piece portions, each supporting piece portion being lowered toward the bottom surface via a contact surface depending from the upper surface to receive both ends of the rectangular preparation. A stepped portion is formed that is partitioned by both side surfaces that stand opposite to each other across the bottom surface. The distance between the both side surfaces is substantially the same as the width dimension of the both end portions for positioning the rectangular slide in the width direction of the both end portions, and the distance between both contact surfaces is longer than the longitudinal dimension of the rectangular preparation. One of the support piece portions is in contact with the end surface of the rectangular preparation placed when protruding from the contact surface through the interior and the other contact surface. At least two buffer screws sandwiching the rectangular preparation in between are attached in such a manner as to be separated from each other in the longitudinal direction of the support piece portion so as to advance and retreat.

According to the apparatus of claim 6, in the fixing member in any one of the apparatuses of claims 1 to 5, the fixing portion partially constituting the fixing member is directly fixed to the movable stage, and thereby the rectangular frame is indirectly fixed. The rectangular preparation is placed on a rectangular frame. At this time, both end portions of the rectangular preparation are placed on the bottom surfaces of the step portions formed on the support piece portions. The width direction of the end portion of the placed rectangular preparation is positioned by being sandwiched between both side surfaces, and the longitudinal direction is similarly positioned by one abutting surface and a buffer screw. Positioning in the width direction is performed at the same time as placing, and positioning in the longitudinal direction is performed when the buffer screw is advanced and brought into contact with the end face of the rectangular preparation. The buffer screw has a function to relieve the inertia caused by stopping and moving the movable stage. The buffer screw is made of a softer material than the slide (If it is too hard, the slide may be damaged). When the screw is tightened, it will be deformed appropriately and between the slide and the wall, between the slide and the screw, and between the screw and the screw hole. It has the effect of facilitating adjustment of the frictional force (vertical drag) between the wall surfaces. (Second problem).

(Feature of the invention of claim 7)
In a tracking device according to a seventh aspect (hereinafter referred to as “the device according to the seventh aspect” as appropriate), as a preferred aspect of the device according to the sixth aspect, the screw member is made of a synthetic resin material.

According to the apparatus of claim 7, since the buffer screw of the apparatus of claim 6 is made of synthetic resin, it is lightweight and easy to manufacture. The degree of the buffering effect can be easily adjusted by appropriately selecting the synthetic resin material.

(Characteristics of the invention described in claim 8)
In the tracking device according to claim 8 (hereinafter referred to as “device of claim 8” as appropriate), as a preferred aspect of the device according to any of claims 1 to 7, the movable stage is installed on a manual stage. The movable stage can be moved by moving the manual stage.

According to the apparatus of claim 8, the movable stage of the apparatus of any one of claims 1 to 7 can be moved by manually moving the manual stage. Since the movable stage and the rectangular preparation move together, the rectangular preparation can be moved indirectly by moving the manual stage. In other words, it is possible to create a pseudo-moving object so that a non-moving object held by a rectangular preparation moves. For this reason, it is not necessary to prepare a living observation target in the development and adjustment of the tracking device, and the labor and cost can be reduced accordingly, and the development and adjustment are facilitated (third problem). .

(Feature of the invention of claim 9)
In the tracking device according to claim 9 (hereinafter, referred to as “device of claim 9” as appropriate), as a preferred aspect of the device of claim 6, the control device is configured to be able to stop tracking in a reversible manner.

According to the device of claim 9, since the control device of the device of claim 8 can be stopped for tracking, first, when it is desired to stop for some reason, secondly, manual tracking is substituted for tracking by the tracking device. This is very convenient when you want to change the tracking target. In other words, if the manual stage is moved while the movement of the movable stage is stopped, tracking as intended by the observer can be performed (fourth problem). If you want to restart after stopping, you can cancel tracking stop and return.

(Features of the invention of claim 10)
In the tracking device according to claim 10 (hereinafter, referred to as “device of claim 10” as appropriate), as a preferable aspect of the device according to any of claims 1 to 9, the objective lens includes a variable focus lens. is there.

According to the apparatus of the tenth aspect, since the objective lens of the apparatus according to any one of the first to ninth aspects is a variable focus lens, it is possible to easily track the movement of the moving object in the Z-axis direction (vertical direction). Can do. The movement in the Z-axis direction includes the movement of the moving animal in the same direction and the vertical vibration of the rectangular preparation, but in any case, a clearer image can be obtained compared to the fixed focus lens. The movement in the Z-axis direction in the three-dimensional tracking can also be omitted (fifth problem).

(Characteristic of the invention of claim 11)
A tracking microscope according to an eleventh aspect (hereinafter, appropriately referred to as a “microscope according to a twelfth aspect”) includes the tracking device according to any one of the first to tenth aspects.

According to the tracking microscope of the eleventh aspect, since the tracking apparatus according to any one of the first to eleventh aspects is included, it is possible to obtain a microscope that makes the best use of the characteristics of each apparatus. That is, it is difficult to generate a tracking failure or a blurred image, it is easy to develop and adjust, the object to be observed can be easily changed, and three-dimensional tracking can be performed without complication.

(Feature of the invention of claim 12)
The tracking microscope according to claim 12 (hereinafter, appropriately referred to as “method of claim 12”) performs two-dimensional imaging of a moving object captured by performing visual feedback on an image of the moving object held on the slide. This is a tracking method for tracking in a three-dimensional manner. Specifically, for an imaging target that continues to be imaged over a predetermined time, a detection procedure for detecting individual relative movements with respect to the slide, and an imaging target for which relative movement has been detected in the detection procedure. A target extraction procedure for extracting as a tracking target candidate, and a selection procedure for selecting an imaging target selected by the user from the tracking target candidates extracted by the target extractor and displayed on the display means, as a tracking target. Have.

According to the method of the twelfth aspect, the moving objects (objects to be imaged) held in the preparation are continuously imaged, and the average relative speed with respect to those preparations is calculated. If the relative velocity with respect to the preparation is zero, the object does not move with respect to the preparation, so it is determined that it is not a moving object and is excluded from the tracking target. The user selects the tracking target while looking at the display means. A captured image usually includes a non-moving object as well as a large number of moving objects, and a tracking target is selected from them. This selection can eliminate non-moving objects and unwanted moving objects. By this elimination, it is possible to effectively eliminate a tracking failure that would otherwise have occurred (first problem).

(Feature of the invention of claim 13)
A tracking microscope according to a thirteenth aspect (hereinafter referred to as “the method of the thirteenth aspect” as appropriate) indicates the coordinates of each imaging target as a tracking target candidate in the target extraction procedure on the premise of the method of the twelfth aspect. The target coordinates are grasped, and in the selection procedure, the point coordinates indicating the coordinates of the point selected by the user on the display means are grasped, and when the point coordinates are grasped, the object related to the coordinates closest to the point coordinates is obtained. An imaging target is selected as a tracking target.

According to the method of the thirteenth aspect, in addition to the operational effect of the method of the twelfth aspect, in the selection procedure, when the coordinates of each of the imaging target as the tracking target candidates are compared with the point coordinates and the point coordinates are grasped. The imaging target related to the coordinates closest to the point coordinates is selected as the tracking target. Each target to be imaged that is a tracking candidate moves relative to the preparation, so that the user's selection may not always be easy. Although the user tries to select a target to be imaged, the timing may not match. In such a case, tracking failure can be effectively suppressed by setting the imaging target closest to the point coordinates as the tracking target (first problem). Used when an imaging target that is different from the target to be selected by the user becomes a tracking target, that is, when an imaging target that is different from the target to be selected is closer to the selection point The person can continue tracking as it is, but can also track the imaging target to be selected by redoing the selection operation.

(Feature of the invention of claim 14)
A tracking method according to claim 14 (hereinafter referred to as “method of claim 14” as appropriate) is a two-dimensional representation of a moving object imaged by performing visual feedback on a captured image of the moving object held on the slide. This is a tracking method for tracking in a three-dimensional manner. Specifically, tracking is performed through a calculation procedure, a determination procedure, an object extraction, and a selection procedure. The calculation procedure is a procedure for calculating an average relative speed with respect to the preparation for an object to be imaged that continues to be imaged over a predetermined time. The determination procedure is a procedure for determining the slow speed by comparing the average relative speed of the imaging target calculated by the calculation procedure with a predetermined reference speed. The target extraction procedure is a procedure for extracting, as a tracking target candidate, the imaged target for which the average relative speed is determined to be faster than the reference speed by the determination unit. Finally, the selection procedure is a procedure for selecting, as a tracking target, an imaging target that satisfies a user's selection command or a predetermined selection condition from among the tracking target candidates extracted by the target extractor.

According to the method of the fourteenth aspect, the moving objects (objects to be imaged) held in the preparation are continuously imaged, and the average relative velocity with respect to those preparations is calculated. If the average relative speed is zero, it is not moving with respect to the slide (slower than the reference speed), so it is determined that it is not a moving object and is excluded from the tracking target. Furthermore, the imaging target whose average relative speed is lower than the reference speed is also excluded from the tracking target. Since what remains to be imaged exceeds the reference speed, these are extracted, and tracking is performed using a selection command from the user or satisfaction of a predetermined selection condition as a tracking target.

According to the present invention, a tracking device and a tracking microscope that do not easily generate a tracking failure or a blurred image, are easy to develop and adjust, can easily change an observation target, and can perform three-dimensional tracking without being complicated. As well as a tracking method.

It is a front view of a tracking microscope. It is a block diagram which shows the structure of a control apparatus. It is a figure which shows a tracking candidate list. It is a perspective view of a fixing mechanism. It is a top view of a fixing mechanism. It is a front view of the fixing mechanism shown in FIG. It is a flowchart which shows operation | movement of a tracking microscope. It is a longitudinal cross-sectional view of a variable focus lens. It is the elements on larger scale of the longitudinal cross-sectional view shown in FIG. It is a flowchart which shows the manufacturing method of a variable focus lens.

Hereinafter, the best mode for carrying out the present invention (hereinafter referred to as “the present embodiment” as appropriate) will be described.

(Tracking microscope structure)
FIG. 1 shows a tracking microscope (tracking microscope, culture microscope) provided with a tracking device according to the present embodiment. The tracking microscope 1 (hereinafter simply referred to as “microscope 1”) is generally composed of a microscope body 3 and a tracking device 21. The microscope main body 3 includes a leg portion 5 positioned at the lowermost end, an inverted L-shaped arm portion 7 that rises from the upper portion of the leg portion 5 and bends at the upper end portion and protrudes forward (frontward in FIG. 1), The lamp 9 is a light source, and a condensing lens 11 that condenses the light from the lamp 9. The tracking device 21 will be described in the next section.

(Tracking device structure)
The tracking device 21 is generally composed of an objective lens 23, an imaging device 25, a movable stage 31, a manual stage 51, and a control device 81. The objective lens 23 is of a revolver type so that a plurality of lenses having different magnifications can be exchanged, and the objective lens 23 is positioned above the condenser lens 11 (at a position facing the object to be imaged). At the bottom. The imaging device 25 is for capturing an image formed through the objective lens 23, and is attached to the top of the arm unit 7. For example, a CMOS imager called “profile imager (trademark)” (manufactured by Hamamatsu Photonics) or the like is suitable for the imaging device 25. The maximum frame rate of the CMOS imager is 2,421 frames / s. An image picked up by the image pickup device 25 is digitally converted by a 12-bit AD converter and sent to the control device 81 as an 8-bit image. For monitoring and acquisition of image data, simultaneous imaging with a high-speed camera may be performed.

The movable stage 31 employs an XY stage capable of two-dimensional movement of the stage part. The movable stage 31 includes a stage unit 33 and a drive unit 35 that drives the stage unit 33, and the drive unit 35 is controlled by a personal computer that functions as a control device 61. The movable stage 31 is entirely installed on the manual stage 51, and the movable stage 31 itself can be moved two-dimensionally by manually controlling the manual stage 51.

(Control device structure)
The structure of the control device will be described with reference to FIG. The control device 61 includes an image input unit 63, an image preprocessing unit 65, an image storage unit 67, an average relative speed calculation unit 69, a speed determination unit 71, a tracking candidate list generation unit 73, a tracking target selection unit 75, and a control command unit 77. And a data storage unit 79 and a stage control unit 81. The image input unit 63 has a function of capturing image data captured by the imaging device 25 and inputting the image data to the image preprocessing unit 65. Note that the control device 61 of the present embodiment may be realized by starting various software that operates on a general-purpose personal computer (not shown) in addition to the hardware described above.

The image preprocessing unit 63 removes fixed pattern noise from the captured image, and the image after the removal is stored in the image storage unit 67. On the other hand, the image after noise removal is sent to the average relative speed calculation unit 69. The average relative speed calculation unit 69 functions as a calculation unit that calculates an average relative speed with respect to the preparation for an object to be imaged (existing in an imaging range for a certain period of time) that has been captured for a predetermined time. The calculated average relative speed data is stored in the data storage unit 79 for each different object to be imaged and is sent to the speed determination unit 71 which is a determination unit. The speed determination unit 71 determines the slow speed of the average relative speed of the object with respect to the reference speed as compared with the reference speed (preset by the user) stored in the data storage unit 79. A group of target data (target data) for which the speed determination unit 71 determines that the average relative speed is faster than the reference speed is extracted, and these data are used as a tracking candidate list generation unit 73 that functions as a target extraction unit. Sent to. The tracking candidate list generation unit 73 writes the sent target data group in the tracking candidate list 85 (see FIG. 3) in association with the target code so as to be searchable. In the present embodiment, the above processing is repeated at a rate of 1000 times per second. In the tracking candidate list 85, for each of the extracted imaging targets, a target code for identifying the target, coordinates indicating the target position with reference to the preparation 15, and a ratio indicating how fast the reference speed is. It is advisable to indicate at least three of the reference speeds. The tracking candidate list 85 is stored in the data storage unit 79 and displayed on the display device 27 so that the user can see it.

The data of the imaging target whose average relative speed is slower than the reference speed is deleted after being stored in the data storage unit 79 and is not sent to the tracking candidate list generation unit 73. Here, the reference speed is set, and the average relative speed is determined by comparing with the reference speed. The fact that the average relative speed is faster than the reference speed indicates the activity, and the activity This is because a certain moving object is considered to be healthy and most suitable for the object to be imaged. Objects to be imaged below the reference speed include both moving objects that are moving but not active, and non-moving objects that do not move at all (for example, microorganisms attached to dust and slides). Is not suitable for tracking, so it is removed from the tracking candidates.

The tracking target selection unit 75 serving as a selection unit receives a selection command input via the input unit 29, selects a target to be imaged corresponding to the selection command from tracking candidates, and selects the control command unit 77 for the selection. Communicate the subject. The control command unit 77 that has received the transmission transmits a control command corresponding to the received transmission to the stage control unit 81. The selection command is performed by the user operating the input unit 29 while looking at the display device 27. In this embodiment, the selection by the user's operation is referred to as selection in the manual tracking mode. On the other hand, in this embodiment, an automatic tracking mode corresponding to the manual tracking mode is also provided. Switching between the manual tracking mode and the automatic tracking mode is performed via the input unit 29. That is, the input unit 29 also serves as a mode switching unit.

When the automatic tracking mode is switched, the tracking target selection unit 75 reads out the selection conditions (preset by the user) stored in the data storage unit 79, and stores the selection conditions in the tracking candidate list 85. The tracking target is selected by comparing with the enumerated speed data. For example, a moving object having a maximum specific reference speed or a moving object that moves at a speed equal to or lower than a reference maximum speed determined separately from the reference speed can satisfy the selection condition.

The object to be imaged (tracking object) selected in the manual tracking mode or the automatic tracking mode is imaged, and the imaged moving object is tracked two-dimensionally or three-dimensionally by performing visual feedback on the image. . In order to realize this, the stage control unit 81 controls the drive unit 35 of the movable stage 31. Note that the control command unit 77 transmits a command to the stage control unit 81 as described above, and manages state transitions according to inputs via the input unit 29.

(Preparate fixture structure)
The fixing mechanism 37 for fixing the preparation 15 to the movable stage 31 will be described with reference to FIGS. The fixing mechanism 37 includes a fixing portion 39 that is directly fixed to the movable stage 31 and a rectangular frame 41 that is supported by the fixing portion 39. Since the fixing mechanism 37 employed in the present embodiment is formed by cutting an aluminum plate of 118 mm × 54 mm × 4 mm, the fixing portion 39 and the rectangular frame 41 are integrated from the time of manufacture, but are manufactured separately. What was integrated later by fixing both by some fixing means can also be used as the fixing mechanism 37. The dimension of the aluminum plate is 72 mm × 22 mm ×? ? Of course, the dimensions are selected in order to provide sufficient margin to hold them with reference to the dimensions of a typical preparation of mm, but it goes without saying that different dimensions can be employed as long as the object of the present invention can be achieved. Yes. Similarly, a plate made of other materials such as a metal plate other than an aluminum plate and a synthetic resin plate, a plate formed by combining different materials such as a metal and a synthetic resin, and other than a plate shape Shaped members can also be used.

Return to the description of the fixing unit 39. A plurality of (four in this case)

screw holes

39h, 39h,. . Through the fixing

screws

39n, 39n,. . The entire fixing mechanism 37 can be fixed to the movable stage 31 by (see FIG. 4). Although the ratio is not limited at all, the fixing portion 39 according to the present embodiment occupies approximately two-thirds of the length of the fixing mechanism 37 in the longitudinal direction (left-right direction in FIG. 5), and the remaining two-thirds are It is occupied by a rectangular frame 41. The rectangular frame 41 includes a support piece portion 42 located on the open end side, a support piece portion 43 that is opposed to the support piece portion 42, and a side piece portion that connects the support piece portion 42 and the support piece portion 43. 46 and 46 form four rectangular sides. The longitudinal dimension of the rectangular opening 41h surrounded by the four sides is formed shorter than the longitudinal dimension L (see FIG. 4) of the preparation 15, and the width dimension is also formed longer than the width dimension W. The reason why the longitudinal dimension is shortened is that, as shown in FIG. 4, both ends of the preparation 15 are spanned and supported below by

step portions

44 and 45 described later. The main reason why the width dimension is long is to secure the width dimension for forming the

step portions

44 and 45.

A step 44 is formed at the approximate center in the longitudinal direction (vertical direction in FIG. 5) of the support piece 42. The step portion 44 opens in the direction of the rectangular opening 41h, and descends toward the bottom surface 42c via a contact surface 42b that hangs down from the upper surface 42a of the support piece portion 42, while both side surfaces face each other across the bottom surface 42c. It is partitioned by 42d and 42d (see FIGS. 5 and 6). The distance L1 between the side surfaces 42d and 42d shown in FIG. 5 is set to a dimension (L1 ≧ W) that is substantially the same as or slightly longer than the width dimension W at both ends of the preparation 15. This is because the both ends of the slide 15 placed on the bottom surface 42c are received without pinching or play so that positioning in the width direction can be easily performed. The distance L2 between the contact surfaces 42b and 43b is set to a dimension (L2> L) longer than the longitudinal dimension L of the preparation.

On the other hand, a step 45 is formed at the approximate center in the longitudinal direction (vertical direction in FIG. 5) of the support piece 43 facing the support piece 42. Similarly to the step portion 44, the step portion 45 also opens in the direction of the rectangular opening 41h. The step portion 45 descends toward the bottom surface 43c via a contact surface 43b depending from the upper surface 43a of the support piece portion 43, and sandwiches the bottom surface 43c. It is partitioned off by

opposite side surfaces

43d and 43d (see FIGS. 5 and 6). As shown in FIG. 5, the distance between both

side surfaces

43d and 43d is set to a dimension (L1 ≧ W) that is substantially the same as or slightly longer than the width W at both ends of the preparation 15, that is, a dimension equal to L1. . This is because, similarly to the stepped portion 44, it is possible to easily perform positioning in the width direction of both end portions of the preparation 15 placed on the bottom surface 43c. As described above, the distance L2 between the contact surfaces 42b and 43b is set to a dimension (L2> L) that is longer than the longitudinal dimension L of the slide. However, this surplus dimension is a buffer screw that will be described later. This is to allow the protrusion of.

Screw holes 42h and 42h are formed through the support piece 42 in the width direction. The screw holes 42h and 42h are holes for allowing the buffer screws 48 and 48 to penetrate the inside of the support piece 42 and protrude from the contact surface 42b so as to be able to advance and retract. The buffer screws 48, 48 are for abutting against the end surface of the rectangular preparation 15 placed on the bottom surface 42c when projecting, and sandwiching the rectangular preparation 15 with the other abutting surface 43b. . At least two buffer screws are necessary, and they are attached with a predetermined distance in the longitudinal direction of the support piece 42. The buffer screws 48, 48 have a function to soften the inertia accompanying the start / stop of the movable stage 31 and the direction change by elastic deformation thereof. Furthermore, the buffer screws 48 also have a function of preventing displacement in the thickness direction of the rectangular preparation 15 by frictional force due to contact with the end surface. These functions generally suppress the vertical vibration of the rectangular preparation so as not to exceed the depth of field of the objective lens. In order to efficiently exhibit such a function as a buffer member, it is preferable that the buffer screws 48 are formed of a synthetic resin material. As shown in FIGS. 4 and 5, a cutout 43 c is formed on the side of the support piece 43. The notch 43c is for inserting the buffer screw 49 from the lateral direction, and the tip of the inserted buffer screw 49 protrudes through the screw hole 43h so as to be able to advance and retreat. The protrusion of the buffer screw 49 is for abutting the tip of the buffer screw 49 to the end surface of the prepared slide 15 and preventing the displacement of the rectangular slide 15 in the thickness direction due to the frictional force caused by the contact. Although the buffer screw 49 is not necessarily required and can be omitted, a function of assisting the function of the buffer screws 48 can be expected by providing the buffer screw 49. In addition, provision of a buffer screw other than the buffer screw 49 is not prevented. Although not shown, it is conceivable to form a portion that covers the upper surface of the preparation 15 and to provide a buffer screw that penetrates the portion 15 and comes into contact with the upper surface.

(Tracking device operation procedure)
The operation procedure (tracking method) of the tracking device will be described with reference to FIGS. Image data captured by the imaging device 25 is captured by the image input unit 63 (S1) and preprocessed by the image preprocessing unit 65 (S3). The preprocessed image data is sent to the image storage unit 67 and sent to the average relative speed calculation unit 69 (S4). The average relative speed calculation unit 69 continuously captures images from the sent image data for a predetermined time. The imaging target being identified is specified (S5). That is, those that initially existed but went out of the imaging range in the middle, and those that entered the imaging range from a certain point in time but have not passed the predetermined time are not specified. With respect to the imaging target thus identified, the average relative speed calculation unit 69 detects the average relative speed for each of the imaging targets (detection procedure) and sends it to the data storage unit 79 and the speed determination unit (S7). . The speed determination unit 71 reads the reference speed from the data storage unit 79 (S9), compares the read reference speed with each of the sent average relative speeds, and determines their slow speed. Here, an imaging target whose average relative speed is faster than the reference speed is set as a tracking target candidate, and only image data related thereto is extracted (target extraction procedure), and the position coordinates (this is referred to as “target coordinates” "). The data is sent to the data storage unit 79 and the tracking candidate list generation unit 73 together with the target coordinate and the specific reference speed indicating how much faster the reference speed is (S11). That is, the imaging target related to the average relative speed (including those in the stopped state) slower than the reference speed is excluded here.

The tracking candidate list generating unit 73 generates a tracking candidate list 85 (see FIG. 3) based on the image data, position coordinates, and ratio reference speed sent from the speed determining unit 71 (S13). The data of the created tracking candidate list 85 is stored in the data storage unit 79 (S14) and simultaneously displayed on the display device 27 (S15). The display on the display device 27 is to enable the user to specify the imaging target.

The tracking target selection unit 75 checks whether the manual tracking mode or the automatic tracking mode is selected, and proceeds to S19 when the manual tracking mode is switched to, and proceeds to S41 when the automatic tracking mode is switched (S17). ). In S19, when there is a selection command waiting for the selection command, the imaging target related to the selection command is selected (selection procedure) and transmitted to the tracking control unit 77 (S21). The tracking control unit 77 that has received the transmission sends a command for tracking the selected imaging target to the movable stage 31 (drive unit 35), and performs two-dimensional tracking by visual feedback (S23). In the above selection procedure, the point coordinates indicating the coordinates of the point selected by the user on the display unit 27 functioning as the display means are grasped, and when the point coordinates are grasped, the object to be imaged relating to the coordinates closest to the point coordinates is obtained. Select target as tracking target. Unless the object to be imaged is lost during tracking, the tracking is continued as it is (S25), and the tracking is ended when an end instruction is received (S27). The termination instruction is based on a termination command from the input unit 29. Returning to S25, if the object to be imaged is lost during tracking, the process returns to S19 and waits for selection of a new object to be imaged.

Returning to S <b> 17, the tracking target selection unit 75 when the automatic tracking mode is switched reads the selection condition from the data storage unit 79 (S <b> 41), and each tracking candidate listed in the tracking candidate list is the read selection condition. The tracking target optimum for the selection condition is selected in light of the specific reference speed and transmitted to the tracking control unit 77 (S43). The tracking control unit 77 continues tracking as long as the subject to be imaged is not lost during tracking (S45, S47, and ends the tracking when receiving the termination instruction (S27). The termination instruction is sent from the input unit 29. When the process returns to S47 and the object to be imaged is lost during tracking, the process returns to S41 and waits for selection of a new object to be imaged. Although two-dimensional tracking is assumed, it goes without saying that three-dimensional tracking may be used.

(Modification of this embodiment)
A modification of the present embodiment (hereinafter referred to as “this modification”) will be described with reference to FIGS. In this modification, a modification of the objective lens 23 of the present embodiment will be described with reference to FIGS.

The variable focus lens 91 includes an accommodating portion 94, a driving portion 101, a first medium 103, a second medium 104, a control portion 105, and a driving stage 106. The accommodating portion 94 includes an accommodating space 95, a first wall portion 96, a second wall portion 13, and a partition portion 98.

The accommodating space 95 is disposed between the first wall portion 96 and the second wall portion 13. Furthermore, the first wall portion 96 and the second wall portion 13 are arranged so as to face each other with the accommodation space 95 interposed therebetween.

The first wall portion 96 includes a first window portion 96a that transmits light. The 2nd wall part 13 is provided with the 2nd window part 97a which further permeate | transmits the light which permeate | transmitted the 1st window part 96a. The first window portion 96a and the second window portion 97a can be configured, for example, by forming through holes in the first wall portion 96 and the second wall portion 13 and then inserting transparent glass into these portions.

The partition part 98 is configured to form a first space 95a and a second space 95b by partitioning the accommodation space 95. The partition part 98 is configured in a flat plate shape in the present modification. The partition portion 98 includes a first surface 98a and a through hole 98b. In the present modification, the upper surface (see FIG. 8) of the partition portion 98 constitutes the first surface 98a. Thereby, the 1st surface 98a comprises at least one part in the surface of the partition part 98. As shown in FIG.

The first surface 98a is arranged in one virtual flat surface. That is, the first surface 98a has a shape that forms part of one virtual flat surface. In FIG. 8, the first surface 98 a coincides with the entire top surface of the partition portion 98. However, as described later, the first surface 98a only needs to exist around the through hole 98b, and the area thereof may be very small.

The through hole 98b is formed so as to penetrate the first surface 98a and penetrate the partition portion 98. The through hole 98b is formed in a cylindrical shape that is circular in cross section. The magnitude | size of the through-hole 98b can be set suitably according to a use. In the present embodiment, the through hole 98b is assumed to have a diameter of about 3.000 ± 0.0001 mm and a processing error, but this is merely an example. In addition, although it is possible to form a large number of through holes 98b in an array, in this modification, one through hole 98b will be described.

The through hole 98b is disposed on the optical path where the light transmitted through the first window 96a is directed to the second window 97a (see FIG. 8). The through hole 98b is configured to transmit the first window portion 96a. A peripheral edge 98c constituting the open end of the through hole 98b is disposed at the end of the first surface 98a. Specifically, in the present modification, the peripheral portion 98c is configured by a line of intersection between the first surface 98a and the inner peripheral surface of the through hole 98b. The shape of the peripheral portion 98c is generally preferably a perfect circle, but in principle, for example, an elliptical shape can be used according to the required shape of the lens.

The first medium 103 and the second medium 104 are made of materials that do not mix with each other in the contact state. The refractive indexes of the first medium 103 and the second medium 104 are different from each other. A combination of such media can be selected as appropriate. For example, a combination of PDMS (Poly-Dimethyl-Siloxane) and pure water can be used. The refractive indices are 1.40 and 1.33, respectively. Either of them may be the first medium 103.

In the description of this specification, it is assumed that the refractive index (n2) of the second medium 104 is smaller than the refractive index (n1) of the first medium 103 (that is, n1> n2) unless otherwise specified. However, it is possible to reverse the magnitude relationship of the refractive index.

In this example, a liquid is used as the first medium 103 and the second medium 104. However, these media can be in the form of a sol, a gel, or an elastic body in addition to the liquid. In short, any medium can be used as long as the change in the pressure received from the drive unit 101 can be applied to the interface between the two media to change the shape of the interface.

The first medium 103 is accommodated in the first space 95a. Similarly, the second medium 104 is accommodated in the second space 95b. Of course, the second medium 104 can be accommodated in the first space 95a. However, for convenience of explanation, in this specification, the medium accommodated in the first space 95a is named the first medium.

In the state of being accommodated in the accommodating space 95, the first medium 103 and the second medium 104 are in contact with each other. The outer periphery of the interface 107 between the first medium 103 and the second medium 104 in this contact state is located at the peripheral edge 98c in the through hole 98b (see FIG. 3).

Further, the density of the first medium 103 and the density of the second medium 104 are substantially equal. The drive unit 101 is configured to change the curvature of the interface 107 by changing the pressure or volume of the first medium 103 or the second medium 104.

In the present modification, the drive unit 101 is mainly composed of a piezo element 102 having a laminated structure. A piezo element is a piezoelectric element that changes its shape in accordance with an applied voltage. By applying an AC voltage, the shape of the piezo element can be periodically expanded and contracted. Since the configuration of the driving unit 101 may be the same as that shown in Non-Patent Documents 6 and 7 described above, detailed description thereof is omitted.

In this modification, the drive unit 101 is configured to apply periodic pressure fluctuations to the first medium 103. Of course, it is also possible to apply a pressure variation to the second medium 104.

The control unit 105 is configured to control the operation of the drive unit 101 by sending a drive signal to the drive unit 101. The drive signal in the control unit 105 may be generated according to previously input data, or may be dynamically generated based on analysis of an image obtained by the lens.

The drive stage 106 is a mechanism for adjusting the position or posture of the drive unit 101 with respect to the first medium 103. Such a drive stage 106 can be easily configured by, for example, a table for fixing the drive unit 101 and an actuator (none of which is shown) that can position the table in a three-dimensional direction, and thus detailed description thereof is omitted. To do.

(Manufacturing method of variable focus lens of this embodiment)
Next, an outline of a method for manufacturing the variable focus lens 91 according to the present embodiment will be described based on the flowchart shown in FIG.

(Step SA-1)
First, a flat partition 98 is prepared. Next, the surface of the partition 98 is processed into a flat surface. Higher surface flatness is preferred. As a method of increasing the flatness, for example, there is a polishing process used when manufacturing a semiconductor tomb board. That is, since a method for increasing the flatness is established in a technique such as a semiconductor manufacturing process, high flatness can be achieved relatively easily by using it.

In the present modification, the flat surface formed in this way can be used as the first surface 98a of the partition portion 98. In this modification, since the first surface 98a is a flat surface, the first surface 98a with high processing accuracy (that is, high flatness) can be obtained relatively easily.

(Step SA-2)
Next, a hole is made in the partition portion 98 to form a through hole 98b. As a specific method of drilling, for example, there is an etching process using photolithography. An example of this processing method is shown below.

In this method, first, a mask portion (for example, SiO ₂ film) is formed on the surface of a substrate (for example, Si substrate) (not shown). Next, after applying a photoresist (photosensitive agent) to the surface of the mask portion, the photoresist is exposed in a predetermined pattern. Thereafter, the exposed photoresist (or the photoresist not exposed to light) is removed. Thereafter, the mask portion is removed by etching based on the pattern transferred to the photoresist. Further, the substrate in the portion where the mask portion is removed is removed by etching. In this way, the through hole 98b can be formed.

In the present embodiment, the first surface 98a is formed with respect to the partition portion 98, and the through hole 98b is formed so as to penetrate the first surface 98a. For this reason, in this technique, in order to form the through-hole 98b, the technique which can obtain high processing precision like photolithography is applicable. That is, according to the present embodiment, the processing accuracy of the through hole 98b can be made extremely high.

In the conventional technology (Non-Patent Documents 6 and 7), as described above, the vicinity of the peripheral edge of the through hole is an inclined surface (conical surface). For this reason, the conventional technique has a problem that it is difficult to form a through-hole using a MEMS technique such as photolithography, and the processing accuracy of the peripheral edge of the through-hole tends to be low. Since the dimensional error in the shape of the peripheral portion of the through hole affects the accuracy of the obtained lens surface, it is difficult to improve the resolution with the conventional lens.

On the other hand, in the technique of the present embodiment, the flat first surface 98a is formed in the partition portion 98, and the through hole 98b is formed so as to penetrate the partition surface 98. Therefore, the processing accuracy of the through hole 98b is increased. Can do. For this reason, with this technique, it becomes easy to increase the dimensional accuracy of the peripheral portion 98c of the through-hole work 42.

Therefore, according to the variable focus lens 91 of the present embodiment, the lens surface formed by the interface 107 can be formed with high accuracy. For this reason, the variable focus lens 91 has an advantage that the resolution of the lens can be improved.

In view of the operation described above, the first surface 98a only needs to contribute to the peripheral portion 98c. That is, the first surface 98a may be processed into a non-flat shape in a portion that does not contribute to the configuration of the peripheral edge portion 98c. For example, after the first surface 98a is processed flat, it can be processed into other shapes. It is also possible to laminate other members on the first surface 98a.

(Step SA-3)
Next, the partition portion 98 is attached to the housing section 11 in the housing portion 94 (see FIG. 2), thereby forming the first space 95a and the second space 95b. After that, first, the first medium 103 is filled in the first space 95a. Then, the liquid level of the first medium 103 gradually rises and reaches the inside of the through hole 98b. The liquid level further rises inside the through hole 98b and reaches the peripheral edge 98c. The operator observes the optical image reflected by the liquid surface of the first medium 103 while the liquid level rises, particularly after reaching the inside of the through hole 98b.

(Step SA-4)
Then, when the magnification of the optical image reflected by the liquid surface of the first medium 103 changes greatly, it is determined that the liquid surface has reached the peripheral portion 98c. At that time, the filling of the first medium 103 is stopped.

Generally, when the first medium 103 rises inside the through hole 98b, the shape of the liquid surface is defined by a contact angle determined by the inner peripheral surface of the through hole and the first medium 103. For this reason, basically, the optical image observed by reflection on the liquid surface does not change.

However, once the liquid level reaches the peripheral edge 98c, the position of the outer periphery of the liquid surface is fixed to the peripheral edge 98c. When the liquid level rises in this state, the curvature of the liquid level changes greatly. For example, a liquid surface that is concave changes to a convex shape. Then, the magnification of the reflected image on the liquid surface changes greatly and is visually recognized by an observer. Of course, this visual recognition can be performed directly by the naked eye, but when the lens is very small, it can be performed through a microscope.

Therefore, if the magnification of the reflected image is greatly changed and the filling of the first medium 103 is stopped, the position of the outer periphery of the liquid surface can be matched with the peripheral portion 98c. Therefore, also from this point, there is an advantage that the accuracy of the lens shape defined by the peripheral portion 98c can be increased.

(Step SA-5)
Next, the second medium 104 is filled in the second space 95b. Thereby, the second medium 104 can be arranged in contact with the upper part of the first medium 103. In this state, an interface 107 (see FIG. 8) is formed at the contact portion between the first medium 103 and the second medium 104. Then, the variable focus lens can be obtained by attaching the drive unit 101 to the housing unit 94. Since the specific manufacturing method other than the above may be the same as the techniques described in Non-Patent Documents 6 and 7, further detailed description is omitted.

(Operation of variable focus lens)
Next, the operation of the variable focus lens according to this embodiment will be described. In the initial state, the position of the focal point due to the lens action at the interface 107 is represented by the symbol F in FIG.

Next, by driving the drive unit 101, the first medium 103 is pressed to cause the interface 107 to protrude upward. Then, the interface 107 bulges upward in FIG. 2 (indicated by a broken line in FIG. 2) while the periphery of the interface 40 remains in contact with the peripheral portion 98c of the through hole 98b. As a result, the position of the focal point due to the lens action at the interface 107 changes to the position indicated by the symbol F 'in FIG. According to this modification, a variable focus lens can be obtained in this way.

The lens of the present modification has the advantage that the shape of the interface 107, that is, the shape of the lens surface can be made highly accurate, so that a high resolution can be obtained even though it is a variable focus lens.

By releasing the pressure on the first medium 103, the focus position can be returned to the initial state. Of course, the focal length can be appropriately adjusted by adjusting the amount of pressing to the first medium 103.

In this modification, since a piezo element is used as the drive unit 101, the focal length can be changed at a high frequency of about 1 kHz, for example.

In this modification, since the liquid is used as the first medium 103 and the second medium 104, the amount of compression of the medium itself is small. For this reason, the pressure applied to the first medium 103 can be immediately transmitted to the second medium 104. Also from this point, the lens of the present embodiment can achieve a high operating frequency.

Furthermore, in this modification, the density of the first medium 103 and the density of the second medium 104 are substantially matched. If the density of the two media is different, the shape of the interface 107 changes due to the influence of gravity when the posture of the lens changes (for example, when the vertical direction of the lens is reversed). For example, a phenomenon occurs in which the interface shape is deviated and decentered. On the other hand, in this embodiment, since the densities of the two media are matched, there is an advantage that the shape of the interface 107 does not change even in such a case.

DESCRIPTION OF SYMBOLS 1 Tracking microscope 3 Microscope main body 5 Leg part 7 Arm part 9 Lamp 11 Condensing lens 15 Preparation 17 High-speed camera 21 Tracking apparatus 23 Objective lens 25 Imaging apparatus 27 Display part 29 Input part 31 Movable stage 33 Stage part 35 Drive part 37 Fixing mechanism 39 fixed part 39a screw 39h screw hole 41 rectangular frame 41h rectangular opening 42 support piece part 43 support piece part 43c notch 44 step part 45 step part 46 side piece part 48 screw 49 screw 51 manual stage 61 control device 63 image input part 65 image Pre-processing unit 67 Image storage unit 69 Average relative speed calculation unit 71 Speed comparison unit 73 Tracking candidate list generation unit 75 Tracking target selection unit 77 Track Control unit 79 data storage unit 81 stage control unit 85 tracking candidate list 91 variable focus lens 94 storage unit 95 storage space 95a first space 95b second space 96 first wall portion 96a first window portion 97 second wall portion 97a first 2 windows 98 partitioning part 98a first surface 98b through hole 98c peripheral part 101 driving part 102 piezo element 103 first medium 104 second medium 105 control part 106 driving stage 107 interface

Claims

A rectangular preparation that holds a moving object;
A movable stage in which the rectangular preparation is fixed via a fixing mechanism;
An objective lens disposed opposite the rectangular preparation;
An imaging device that captures an image of a moving object imaged by the objective lens;
A display device that visually displays an image of the moving object imaged by the imaging device;
A control device for controlling the movable stage in order to track a moving object imaged two-dimensionally or three-dimensionally by performing visual feedback on the object image imaged by the imaging device;
In a tracking device comprising:
The control device
For the imaging target that continues to be imaged over a predetermined time,
Detection means for detecting individual relative movement with respect to the preparation;
Target extraction means for extracting the imaged target whose relative movement has been detected by the detection means as a tracking target candidate;
A selection means for selecting, as a tracking target, an imaging target selected by a user from among tracking target candidates extracted by the target extraction hand and displayed on the display means;
A tracking device comprising:
The target extracting means is configured to grasp target coordinates indicating coordinates of each of the imaging targets as tracking target candidates,
The selection means is configured to grasp point coordinates indicating the coordinates of the point selected by the user on the display means;
The tracking device according to claim 1, wherein the selection unit is configured to select an object to be imaged according to a coordinate closest to the point coordinate as a tracking target when the point coordinate is grasped. .
A rectangular preparation that holds a moving object;
A movable stage in which the rectangular preparation is fixed via a fixing mechanism;
An objective lens disposed opposite the rectangular preparation;
An imaging device that captures an image of a moving object imaged by the objective lens;
A display device that visually displays an image of the moving object imaged by the imaging device;
A control device for controlling the movable stage in order to track a moving object imaged two-dimensionally or three-dimensionally by performing visual feedback on the object image imaged by the imaging device;
In a tracking device comprising:
The control device
A calculation means for calculating an average relative speed with respect to the preparation for an imaging target that continues to be imaged over a predetermined time;
A determination means for determining a slow speed by comparing an average relative speed of the imaging target calculated by the calculation means with a predetermined reference speed;
Target extraction means for extracting the imaged target, which has been determined by the determination means to have the average relative speed faster than the reference speed, as a tracking target candidate;
A selection means for selecting, as a tracking target, a target to be imaged that satisfies a user's selection command or a predetermined selection condition from among tracking target candidates extracted by the target extractor;
A tracking device comprising:
A rectangular preparation that holds a moving object;
A movable stage in which the rectangular preparation is fixed via a fixing mechanism;
An objective lens disposed opposite the rectangular preparation;
An imaging device that captures an image of a moving object imaged by the objective lens;
A control device for controlling the movable stage in order to track a moving object imaged two-dimensionally or three-dimensionally by performing visual feedback on the object image imaged by the imaging device;
In a tracking device comprising:
The control device
An object designating means for arbitrarily design an imaging target;
Target selection means for selecting the imaging target specified by the target specification means as a tracking target;
A tracking device comprising:
A rectangular preparation that holds a moving object;
A movable stage in which the rectangular preparation is fixed via a fixing mechanism;
An objective lens disposed opposite the rectangular preparation;
An imaging device that captures an image of a moving object imaged by the objective lens;
A control device for controlling the movable stage in order to track a moving object imaged two-dimensionally or three-dimensionally by performing visual feedback on the object image imaged by the imaging device;
In a tracking device comprising:
The control device
A calculation means for calculating an average relative speed with respect to the chamber for an imaging target that continues to be imaged over a predetermined time,
A determination means for determining a slow speed by comparing an average relative speed of the imaging target calculated by the calculation means with a predetermined reference speed;
Target extraction means for extracting the imaged target, which has been determined by the determination means to have the average relative speed faster than the reference speed, as a tracking target candidate;
A selection means for selectively selecting an imaging target according to a user's selection command or satisfaction of a predetermined selection condition from the tracking target candidates extracted by the target extractor;
Mode switching means for switching between manual tracking mode and automatic tracking mode,
When the mode switching unit is switched to the manual tracking mode, the selection unit is selected according to a user's selection command, and when the mode switching unit is switched to the automatic tracking mode, A tracking device configured to select an imaging target as a tracking target according to satisfaction.
The fixing mechanism includes a fixing portion that is directly fixed to the movable stage, and a rectangular frame supported by the fixing portion,
The rectangular frame is configured to include a pair of support pieces for supporting the both ends in the longitudinal direction of the rectangular preparation downward,
Each of the support piece portions has a step partitioned by both side surfaces that are opposed to each other while sandwiching the bottom surface while descending toward the bottom surface via a contact surface that hangs down from the top surface in order to receive both ends of the rectangular preparation. Part is formed,
The distance between the both side surfaces is substantially the same as the width dimension of the both end portions for positioning the rectangular slide in the width direction of the both end portions, and the distance between both contact surfaces is longer than the longitudinal dimension of the rectangular preparation. Set
Either one of the support piece portions is in contact with the end surface of the rectangular preparation placed when protruding from the contact surface through the inside, and the rectangular preparation plate is in contact with the other contact surface. The tracking device according to any one of claims 1 to 5, wherein at least two buffer screws that sandwich the shaft are attached so as to be able to move forward and backward apart from each other in the longitudinal direction of the support piece.
The tracking device according to claim 6, wherein the buffer screw is made of a synthetic resin material.
The movable stage is installed on a manual stage;
The tracking apparatus according to claim 1, wherein the movable stage is configured to be moved by moving the manual stage.
The tracking device according to claim 8, wherein the control device is configured to be able to stop tracking so as to be able to return.
The tracking device according to claim 1, wherein the objective lens includes a variable focus lens.
A tracking microscope comprising the tracking device according to claim 1.
In a tracking method for tracking a moving object imaged by performing visual feedback on a captured image of the moving object held in a preparation,
A detection procedure for detecting an individual relative movement with respect to the preparation for an imaging target that continues to be imaged over a predetermined time,
A target extraction procedure for extracting the imaged target whose relative movement is detected in the detection procedure as a tracking target candidate;
A selection procedure for selecting the imaging target selected by the user from the tracking target candidates extracted by the target extractor and displayed on the display means, as a tracking target;
A tracking method comprising:
In the target extraction procedure, grasp the target coordinates indicating the coordinates of each target to be imaged as tracking target candidates,
In the selection procedure, the point coordinates indicating the coordinates of the point selected by the user on the display means are grasped, and when the point coordinates are grasped, the object to be imaged that is closest to the point coordinates is set as the tracking target. The tracking method according to claim 12, wherein the tracking method is selected.
In a tracking method for tracking a moving object imaged two-dimensionally or three-dimensionally by performing visual feedback on a captured image of the moving object held in a preparation,
A calculation procedure for calculating an average relative speed with respect to the preparation for the imaging target that continues to be imaged over a predetermined time,
A determination procedure for determining the slow speed by comparing the average relative speed of the imaging target calculated in the calculation procedure with a predetermined reference speed;
A target extraction procedure for extracting, as a tracking target candidate, the imaged target for which the determination unit has determined that the average relative speed is faster than the reference speed;
A selection procedure for selecting, as a tracking target, an imaging target corresponding to satisfaction of a user's selection command or a predetermined selection condition from among tracking target candidates extracted by the target extractor;
A tracking method comprising: